Monteiro Ana Carolina, Scovino Aline, Raposo Susane, Gaze Vinicius Mussa, Cruz Catia, Svensjö Erik, Narciso Marcelo Sampaio, Colombo Ana Paula, Pesquero João B, Feres-Filho Eduardo, Nguyen Ky-Anh, Sroka Aneta, Potempa Jan, Scharfstein Julio
Carlos Chagas Filho Institute of Biophysics, Institute of Biomedical Sciences, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
J Immunol. 2009 Sep 15;183(6):3700-11. doi: 10.4049/jimmunol.0900895. Epub 2009 Aug 17.
Porphyromonas gingivalis, a Gram-negative bacterium that causes periodontitis, activates the kinin system via the cysteine protease R-gingipain. Using a model of buccal infection based on P. gingivalis inoculation in the anterior mandibular vestibule, we studied whether kinins released by gingipain may link mucosal inflammation to T cell-dependent immunity through the activation of bradykinin B(2) receptors (B(2)R). Our data show that P. gingivalis W83 (wild type), but not gingipain-deficient mutant or wild-type bacteria pretreated with gingipain inhibitors, elicited buccal edema and gingivitis in BALB/c or C57BL/6 mice. Studies in TLR2(-/-), B(2)R(-/-), and neutrophil-depleted C57BL/6 mice revealed that P. gingivalis induced edema through the sequential activation of TLR2/neutrophils, with the initial plasma leakage being amplified by gingipain-dependent release of vasoactive kinins from plasma-borne kininogens. We then used fimbriae (Fim) Ag as a readout to verify whether activation of the TLR2-->PMN-->B(2)R axis (where PMN is polymorphonuclear neutrophil) at early stages of mucosal infection had impact on adaptive immunity. Analyzes of T cell recall responses indicated that gingipain drives B(2)R-dependent generation of IFN-gamma-producing Fim T cells in submandibular draining lymph nodes of BALB/c and C57BL/6 mice, whereas IL-17-producing Fim T cells were generated only in BALB/c mice. In summary, our studies suggest that two virulence factors, LPS (an atypical TLR2 ligand) and gingipain, forge a trans-cellular cross-talk between TLR2 and B(2)R, thus forming an innate axis that guides the development of Fim-specific T cells in mice challenged intrabuccally by P. gingivalis. Ongoing research may clarify whether kinin-driven modulation of T cell responses may also influence the severity of chronic periodontitis.
牙龈卟啉单胞菌是一种可引发牙周炎的革兰氏阴性菌,它通过半胱氨酸蛋白酶R-牙龈蛋白酶激活激肽系统。我们采用在前下颌前庭接种牙龈卟啉单胞菌的颊部感染模型,研究牙龈蛋白酶释放的激肽是否可通过激活缓激肽B(2)受体(B(2)R)将黏膜炎症与T细胞依赖性免疫联系起来。我们的数据显示,牙龈卟啉单胞菌W83(野生型),而非牙龈蛋白酶缺陷型突变体或经牙龈蛋白酶抑制剂预处理的野生型细菌,可在BALB/c或C57BL/6小鼠中引发颊部水肿和牙龈炎。对TLR2(-/-)、B(2)R(-/-)和中性粒细胞耗竭的C57BL/6小鼠的研究表明,牙龈卟啉单胞菌通过TLR2/中性粒细胞的顺序激活诱导水肿,最初的血浆渗漏通过血浆源性激肽原中血管活性激肽的牙龈蛋白酶依赖性释放而放大。然后,我们使用菌毛(Fim)抗原作为指标,以验证黏膜感染早期TLR2->PMN->B(2)R轴(其中PMN是多形核中性粒细胞)的激活是否对适应性免疫有影响。对T细胞回忆反应的分析表明,牙龈蛋白酶在BALB/c和C57BL/6小鼠的下颌下引流淋巴结中驱动产生IFN-γ的Fim T细胞的B(2)R依赖性生成,而产生IL-17的Fim T细胞仅在BALB/c小鼠中生成。总之,我们的研究表明,两种毒力因子,即脂多糖(一种非典型TLR2配体)和牙龈蛋白酶,在TLR2和B(2)R之间形成了一种跨细胞的相互作用,从而形成了一个先天性轴,该轴指导在颊部受到牙龈卟啉单胞菌攻击的小鼠中Fim特异性T细胞的发育。正在进行的研究可能会阐明激肽驱动的T细胞反应调节是否也会影响慢性牙周炎的严重程度。